WO2017025984A1 - Smartphone integrated real - time molecular diagnostic device - Google Patents
Smartphone integrated real - time molecular diagnostic device Download PDFInfo
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- WO2017025984A1 WO2017025984A1 PCT/IN2016/050263 IN2016050263W WO2017025984A1 WO 2017025984 A1 WO2017025984 A1 WO 2017025984A1 IN 2016050263 W IN2016050263 W IN 2016050263W WO 2017025984 A1 WO2017025984 A1 WO 2017025984A1
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- smart phone
- nucleic acid
- detection
- amplification
- smartphone
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Classifications
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L7/00—Heating or cooling apparatus; Heat insulating devices
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/06—Auxiliary integrated devices, integrated components
- B01L2300/0627—Sensor or part of a sensor is integrated
- B01L2300/0654—Lenses; Optical fibres
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2300/00—Additional constructional details
- B01L2300/18—Means for temperature control
- B01L2300/1805—Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
- B01L2300/1822—Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks using Peltier elements
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/01—Arrangements or apparatus for facilitating the optical investigation
- G01N21/03—Cuvette constructions
- G01N21/0332—Cuvette constructions with temperature control
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2201/00—Features of devices classified in G01N21/00
- G01N2201/02—Mechanical
- G01N2201/022—Casings
- G01N2201/0221—Portable; cableless; compact; hand-held
Definitions
- Figure 1 is a schematic illustration of the structure of the smartphone integrated device for amplification and detection of nucleic acid.
- the device consists of the amplification tray (1) containing the space for the placing the reaction chamber (3) and the detection tray (2) containing the optical components and LED.
- amplification tray can be slide out for placing the reaction chamber.
- the amplification tray and the detection tray were placed one above the other with the smartphone placed on top (4).
- the detection module was kept inverted over the amplification module such that the optical components are aligned in front of the camera of the smartphone (5).
- the assay was performed using the genomic DNA extracted from Staphylococcus auresus and a no template control was used by addition of nuclease free water instead of genomic DNA.
- the amplification was performed on the smart phone device using 5 ⁇ 1 of the above reaction mixture on the polymer micro chamber and overlaid with the 2 ⁇ 1 of mineral oil.
- the microchambers were maintained at 63°C for 30 minutes followed by 82°C for 2 minutes to stop the reaction.
- the Figure 5 represents the screenshot of the actual experiment performed on the smartphone device.
- Fig 5a shows the preview of the image captured at the end of the assay shows the fluorescence signal obtained from the control and test sample detected using the detection module attached to the camera of the smart phone.
Abstract
The current invention consists of a portable handheld device for amplification of nucleic acid followed by detection and processing of the test results on a smart phone. The smartphone interfaced temperature module enables isothermal amplification of nucleic acid in less than one hour. The fluorescence signal from the test sample upon amplification was captured by using a simple fluorescence detection module attached to the in-build camera of the smart phone. Further, the captured images were processed to obtain the intensity profile in real time using an automatic image processing application in the smart phone. The current invention demonstrates the smart phone integrated diagnostic device that can form an end to end solution for multiplex, point of care nucleic acid detection of pathogens. It is hand held, user friendly and rapid compared to conventional real time nucleic acid detection platform. In total, the present invention can be next generation diagnostic platform which provides laboratory quality results in a smart phone.
Description
SMARTPHONE INTEGRATED REAL - TIME MOLECULAR DIAGNOSTIC
DEVICE
FIELD OF THE INVENTION
A hand held device for isothermal amplification cum real time detection of nucleic acid on a smart phone.
BACKGROUND OF THE INVENTION
To make nucleic acid detection under field settings, portable device which can amplify and detect the pathogen under point of care is required. There are numerous features limiting the applicability of PCR in many situations. The approach requires thermal cycling instrumentation, considerable technical expertise, and a substantial amount of space and electricity in routine diagnostic laboratories. This limits the use of nucleic acid diagnosis to sophisticated laboratory facilities. The isothermal amplification is a potential alternative to PCR and can be adapted for field settings by using a simple heat block that can maintain the stable isothermal temperature for amplification. Nowadays smart phone technology contains fast processing CPUs (1.0 GHz), high resolution cameras (8 and 13 megapixel) and flexible developmental platform to customize the application which can be used as nucleic acid diagnostic device in resource limited settings. This combination of integrating the smart phone based amplification cum detection module satisfies the need for point of care nucleic acid detection of pathogens. BRIEF DESCRIPTION OF THE DRAWINGS
A description of various aspects, features and embodiments of the present invention is provided herein with reference to the accompanying drawings. The drawings are illustrative and in wired views which are not necessarily drawn to scale.
Figure 1 is a schematic illustration of the structure of the smartphone integrated device for amplification and detection of nucleic acid. In detail, the device consists of the amplification tray (1) containing the space for the placing the reaction chamber (3) and the detection tray (2) containing the optical components and LED. In operation, amplification
tray can be slide out for placing the reaction chamber. The amplification tray and the detection tray were placed one above the other with the smartphone placed on top (4). The detection module was kept inverted over the amplification module such that the optical components are aligned in front of the camera of the smartphone (5).
Figure 2 is a schematic illustration of the amplification module containing the reaction chamber and the heating unit. The isothermal amplification was performed on the polymer (eg: PDMS, PMMA, polypropylene) microchambers with the reaction mixture overlaid with mineral oil to prevent evaporation. The reaction chamber is preferably designed such that the minimal usage of the reaction volume from few microliters can be achieved. However, the reaction chamber can be of any form, shape and structures other than the particular illustrated. The temperature for isothermal amplification was provided by 15mm x 15mm of Peltier thermoelectric device (6) with a heat block (7) controlled by H-bridge and microcontroller unit using smart phone interface. The temperature can be varied between 0 - 100°C suitable for nucleic acid amplification.
Figure 3 is the 3D printed case used for placing the LED, excitation filter and dichroic. In operation, the detection module was kept inverted over the amplification module such that the optical components are aligned in front of the camera of the smartphone. The smartphone can be placed above the device with back camera fixed with the piano convex lens and emission filter using a 3D printed case. Together the LED, excitation filter, dichroic, planoconvex lens and emission filter attached to the camera of the smartphone forms the detection module. The LED light source of suitable peak wavelength is appropriately positioned to illuminate the fluorophore in the test sample. The intensity of the light source was also controlled by the smartphone interface. The light passes through the filter set consisting of the excitation filter (8), dichroic (9) and the fluorescence signal from the test sample was captured using the in-build camera of the smart phone attached with the emission filter and plano-convex lens. The excitation filter is positioned in front of the LED and the dichroic filter is inclined at an angle of 45° that reflect the excitation light evenly on to the reaction chamber. The fluorescence signal from the test sample is detected after passing through the emission filter and the plano-convex lens attached in front of the smartphone camera.
Figure 4 describes the layout of the device. The device consists of an amplification
tray (1) containing the peltier thermoelectric device (6) with the heat block (7) placed over the heat sink (10). The polymer chamber(3) was placed above the heat block . The heating unit was driven by H-bridge (11) and temperature was monitored using the temperature sensor (12) controlled by microcontroller (13) unit using smart phone interface. The fluorescence detection tray (2) was attached to the camera of the smartphone for detection. The fluorescence detection tray consists of the LED (14), excitation filter (8) and dichroic mirror (9). The intensity of the light source was also controlled by the smartphone interface (4). The light passes through the detection tray and the fluore scence signal from the test sample was captured using the in-build camera of the smart phone placed above the emission filter and piano convex lens (5). The entire device is battery powered (15) and the results can be visualized real time on the smart phone.
Fig (5) and (6) describes the graphical outputs.
OBJECTIVE OF THE INVENTION
This invention uses the smartphone to control the parameters for isothermal amplification and real time detection of nucleic acid using the camera of the smart phone. The smartphone interfaced temperature module enables isothermal amplification of nucleic acid in less than one hour compared to sophisticated PCR machine used for detection. This greatly reduces the cost and time required for molecular diagnosis. The fluorescence signal from the test sample upon amplification was captured by using a simple fluorescence detection module attached to the in-build camera of the smart phone. Thus the smartphone avoids the use of specific photodetector used for detection in conventional systems. Further, the captured images were processed to obtain the intensity profile in real time using an custom developed automatic image processing application in the smart phone. The current invention demonstrates the point of care integrated diagnostic device that can form an end to end solution for nucleic acid detection of pathogens. It is hand held, user friendly and rapid compared to conventional real time nucleic acid detection platform. In total, the present invention can be next generation diagnostic platform which provides laboratory quality results in a smart phone.
SUMMARY OF THE INVENTION
The present invention simplifies the molecular diagnosis by utilizing the smart phone for amplification and detection. In particular, the device uses a simple optical attachment to the camera of the smart phone which is used for capturing the fluorescence images from the test sample. This avoids the cost of the sophisticated detectors used for capturing the fluorescence signal. The automatic image processing application on the smart phone can simultaneously process the multiplex fluorescence signal in real - time to provide the graphical readout. The readout is indicative of the presence or absence of the target nucleic acid in addition to the antibiotic sensitivity/resistance profile. In an embodiment, the temperature required for the isothermal amplification is controlled using the smart phone interface which reduces the cost of additional hardware. The entire device was battery powered, smart phone interfaced, portable to enable field diagnostics and low cost compared to other sophisticated real - time nucleic acid amplification platforms.
EMBODIMENTS OF THE INVENTION
In an embodiment, the present invention provides a smart phone device for isothermal amplification of nucleic acid, wherein said device consists of an amplification tray (1) containing the peltier thermoelectric device (6) with the heat block (7) placed over the heat sink (10), the polymer chamber(3) was placed above the heat block, a heating unit was driven by H-bridge (11) and temperature was monitored using the temperature sensor (12) controlled by a microcontroller (13) unit using smart phone interface, a fluorescence detection tray (2) was attached to the camera of the smartphone for detection, the fluorescence detection tray consists of the LED (14), excitation filter (8) and dichroic mirror (9), intensity of the light source was also controlled by the smartphone interface (4), light passes through the detection tray and the fluorescence signal from the test sample was captured using the in-build camera of the smart phone placed above the emission filter and piano convex lens (5), entire device is battery powered (15) and the results is visualized real time on the smart phone. In another embodiment, the present invention provides a smart phone device for isothermal amplification of nucleic acid, wherein the amplification module is capable of isothermal amplification, PCR, RT-PCR of DNA/RNA for detection and antibiotic resistance profiling achieved in less than one hour.
In yet another embodiment, the present invention provides a smart phone device for isothermal amplification of nucleic acid, wherein the reaction chamber made up of polymers as polymer micro chambers, microfluidics, PCR tubes or custom designed polymer tubes which includes polypropylene, polystyrene, polyethene, etc.
In one another embodiment, the present invention provides a smart phone device for isothermal amplification of nucleic acid, wherein a fluorescence module attached to the camera of the smartphone or other electronic sensor for capturing the images from the micro chambers, a battery powered LED light source to excite the detection probe such as intercalating dye, calcein, hairpin probes, nanoparticles based fluorescence probes etc from the test sample.
In another embodiment, the present invention provides a smart phone device for isothermal amplification of nucleic acid, wherein the captured images were processed in real time on android / iOS/ Windows platform to obtain the fluorescence or colorimetric intensity as a graphical or digital output in the smart phone.
In another embodiment, the present invention provides a smart phone device for isothermal amplification of nucleic acid as claimed in claim 1, wherein an automatic image processing application installed on smart phone for real time simultaneous measuring of the total intensity in the individual reaction microchambers to provide the graphical output of the intensity and the values were in downloadable and convertible Form.
DETAILED DESCRIPTION OF THE INVENTION
The present invention utilizes the rapid amplification of nucleic acid using isothermal amplification and its detection using a simple, portable smartphone device. The isothermal amplification was performed on the polymer (eg: PDMS, PMMA, polypropylene) microchambers with the reaction mixture overlaid with mineral oil to prevent evaporation. The polymer reaction chamber is positioned above the heating block of the amplification module which provides the reaction conditions for the assay. The heating block may be a thermoelectric cooler that maintain the temperature for amplification in the assay chambers. The parameter of the heating block can be provided and controlled or monitored by the smartphone user interface. A microcontroller in the device is provided to control the heating
of the block, monitor the temperature sensors through a H bridge driver using the wireless/ serial/ Bluetooth communication received from the smartphone. The wireless communication electronics is also provided in the device to receive commands from the smartphone. In another embodiment, a light source is positioned inside the detection module to uniformly excite the samples in the assay chambers. The intensity of the light source and commands for on/off status of the LED can be communicated using the smartphone. The LED is turned on at appropriate times by the processing device to excite the sample in the assay when the camera of the smartphone captures the images of the assay. The light source passes through the excitation filter positioned in front of the LED and reflected by the dichroic filter to reach the assay chamber. The fluorescence signal from the assay chambers is recorded as images using the camera of the smartphone attached with piano convex lens and emission filter. The invention is specifically designed to block extraneous light from entering the device, such that interference with optical measurements is reduced or eliminated. The image capturing device comprising the inbuilt camera of the smartphone with the planoconvex lens and emission filter attached to it. The entire device is battery powered and controlled by communication received from the smart phone.
The assay starts with inserting the polymer chamber containing the reaction into the amplification module. Then the application in the smartphone can be initiated to input of reaction parameters for controlling the temperature and LED followed by commands to capture of the images of the assay chamber. At specific time interval the assay chambers are excited and images were captured through the camera of the smartphone over different time interval from few seconds to minutes during amplification from individual reaction chambers (4 or more). The images captured are processed using a custom developed android based automatic image processing application running on the smart phone. The images captured were analyzed and determined to provide the intensity from four or more test sample and displayed as a graphical output with respect to various time interval in the smart phone user interface. The increase in intensity profile upon amplification can be directly correlated to the presence or absence of pathogen in the test sample. The images and the intensity data set can be stored in memory of the smartphone and can be communicated to other devices and database after the end of the assay. The entire device is lithium polymer (3.7V) battery powered and the results can be visualized in real time on the smart phone user interface. The developed smart phone device helps in detection of pathogen in less than an hour under field
conditions. The unique combination turns the smartphone into a device capable of nucleic acid detection within one hour even under remote places, resource limited settings and routine clinical laboratory. This invention overcomes the need for sophisticated real time thermal cycler used for detection.
EXAMPLES
Example 1: LAMP based detection of methiciUin resistant Staphylococcus aureus The LAMP based detection was utilized for differentiation of methiciUin sensitive and resistant strains of Staphylococcus aureus. The primers were designed for the femA gene which is essential for methiciUin resistance and is universally present in all Staphylococcus aureus. In addition, primers were designed targeting the mecA gene which confers resistance to antibiotics. Thus the presence of mecA gene is an indicator for detection of methiciUin resistance Staphylococcus aureus. The crude DNA was prepared from Staphylococcus aureus by boiling the cell suspension at 95°C for 15 minutes followed by freezing at -20°C for 5 minutes. After a brief centrifugation the supernatant containing the crude DNA was directly used for the amplification. The LAMP reaction consist of 12.5 μΐ of 20mM Tris buffer (pH 8.8), 10 mM KC1, 8mM MgS04, lOmM (NH4)2S04, 0.1% Tween -20, 1M Betaine, 1.4mM dNTPs, 0.2μΜ outer primer, 0.8 μΜ loop primer, 1.6 μΜ inner primer, 8U Bst DNA polymerase, 2μ1 of genomic DNA, 1 μΐ of Sybr green dye and rest of nuclease free water to make the final volume of 25μ1. The assay was performed using the genomic DNA extracted from Staphylococcus auresus and a no template control was used by addition of nuclease free water instead of genomic DNA. The amplification was performed on the smart phone device using 5μ1 of the above reaction mixture on the polymer micro chamber and overlaid with the 2μ1 of mineral oil. The microchambers were maintained at 63°C for 30 minutes followed by 82°C for 2 minutes to stop the reaction. The Figure 5 represents the screenshot of the actual experiment performed on the smartphone device. Fig 5a shows the preview of the image captured at the end of the assay shows the fluorescence signal obtained from the control and test sample detected using the detection module attached to the camera of the smart phone. Fig 5b shows the smart phone application (in-house custom developed and not made available in public domain) provides the intensity profile of the test sample over various time intervals in real time as a graphical readout during amplification. The increase in the intensity profile of the test sample indicates the presence of methiciUin resistance genes in the
Staphylococcus aureus. This enables the simultaneous detection of pathogen and determining the antibiotic resistance/sensitivity pattern in less than an hour using a smart phone. Examples of implementations of embodiments of the invention are described above. Modifications may be made to those examples without departing from the spirit and scope of the invention. Fig 5c shows the amplification pattern observed using agarose gel electrophoresis.
Example 2: LAMP based detection of Mycobacterium tuberculosis
The primers for M. tuberculosis complex were designed to target the gyrB gene sequences and amplified using Loop mediated isothermal amplification on the micro chambers of the smart phone device. The genomic DNA was extracted from H37Rv strain of Mycobacterium tuberculosis by phenol-chloroform method and the purified DNA was used for amplification. The LAMP reaction conditions were optimized by varying the MgS04 concentration from 2mM to lOmM. Also the betaine concentration was varied from 0.2M to 1.5M. The LAMP reaction consist of 20mM Tris buffer (pH 8.8), 10 mM KC1, 8mM MgS04, lOmM (NH4)2S04, 0.1% Tween -20, 1M Betaine, 1.4mM dNTPs, 0.2μΜ outer primer, 0.8 μΜ loop primer, 1.6 μΜ inner primer, 8U Bst DNA polymerase, 2μ1 of genomic DNA, Ι μΐ of Sybr green dye and rest of nuclease free water to make the final volume of 25μ1. The amplification was performed 30 - 40 minutes by maintaining the temperature at 63°C. During amplification, the test samples were excited using LED light source and the fluorescence signal upon excitation of Sybr green dye from the test sample was captured using the smart phone camera for every 40 seconds and processed to obtain the real time detection of Mycobacterium tuberculosis for 30 - 40 minutes. Multiple reaction chambers can be simultaneously excited and assayed targeting the drug resistance genes to obtain the multi- drug resistance profile of the pathogen. The Figure 6 represents the screenshot of the actual experiment performed on the smartphone device. Fig 6a shows the preview of the image captured at the end of the assay shows the fluorescence signal obtained from the control and test sample detected using the detection module attached to the camera of the smart phone. Fig 6b shows the smart phone application (in-house custom developed and not made available in public domain) provides the intensity profile of the test sample over various time intervals in real time as a graphical readout during amplification. The increase in the intensity profile of the test sample indicates the presence of Mycobacterium tuberculosis. Fig 6c shows the amplification pattern observed using agarose gel electrophoresis. Examples of implementations of embodiments of the invention are described above. Modifications may be
made to those examples without departing from the spirit and scope of the invention, which is defined in the claims, below.
ADVANTAGES OF THE INVENTION:
The available molecular diagnostic platforms in the market require sophisticated hardware and detection module which increase the cost of diagnosis. Also, the bulky instrumentation cannot be portable and cannot be used in resource limited settings. 1. The present invention is a smart phone integrated device, as it plays an important role in controlling the temperature for amplification, controlling the LED for excitation and smartphone camera is used as detector and smartphone application process the result. The device is not functional without smartphone. 2. The detection was obtained using the simple optical attachment to the smart phone camera and processed using the image processing application of the smartphone.
3. The developed platform is highly portable, battery operated, smart phone integrated temperature control followed by fluorescence detection using smart phone application for real - time multiplex quantitative molecular detection.
4. The invention provides end to end solution starting from amplification to real time multiplex fluorescence detection in less than one hour using smart phone as a molecular diagnostic device.
Claims
A smart phone device for isothermal amplification of nucleic acid, wherein said device consists of an amplification tray (1) containing the peltier thermoelectric device (6) with the heat block (7) placed over the heat sink (10), the polymer chamber(3) was placed above the heat block, a heating unit was driven by H-bridge (11) and temperature was monitored using the temperature sensor (12) controlled by a microcontroller (13) unit using smart phone interface, a fluorescence detection tray (2) was attached to the camera of the smartphone for detection, the fluorescence detection tray consists of the LED (14), excitation filter (8) and dichroic mirror (9), intensity of the light source was also controlled by the smartphone interface (4), light passes through the detection tray and the fluorescence signal from the test sample was captured using the in-build camera of the smart phone placed above the emission filter and piano convex lens (5), entire device is battery powered (15) and the results is visualized real time on the smart phone.
A smart phone device for isothermal amplification of nucleic acid as claimed in claim 1, wherein the amplification module is capable of isothermal amplification, PCR, RT- PCR of DNA/RNA for detection and antibiotic resistance profiling achieved in less than one hour.
A smart phone device for isothermal amplification of nucleic acid as claimed in claim 1, wherein the reaction chamber made up of polymers as polymer micro chambers, microfluidics, PCR tubes or custom designed polymer tubes which includes polypropylene, polystyrene, polyethene, etc.
A smart phone device for isothermal amplification of nucleic acid as claimed in claim 1, wherein a fluorescence module attached to the camera of the smartphone or other electronic sensor for capturing the images from the micro chambers, a battery powered LED light source to excite the detection probe such as intercalating dye, calcein, hairpin probes, nanoparticles based fluorescence probes etc from the test sample.
A smart phone device for isothermal amplification of nucleic acid as claimed in claim 1, wherein the captured images were processed in real time on android / iOS/ Windows platform to obtain the fluorescence or colorimetric intensity as a graphical or digital output in the smart phone.
A smart phone device for isothermal amplification of nucleic acid as claimed in claim 1, wherein an automatic image processing application installed on smart phone for real time simultaneous measuring of the total intensity in the individual reaction microchambers to provide the graphical output of the intensity and the values were in downloadable and convertible Form.
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| IN2425/DEL/2015 | 2015-08-07 |
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| WO2021236987A3 (en) * | 2020-05-20 | 2021-12-23 | The Administrators Of The Tulane Educational Fund | Crispr-based assay for detecting pathogens in samples |
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| ES2939909A1 (en) * | 2022-08-30 | 2023-04-27 | Thenextpangea S L | DEVICE TO DETECT INDUSTRIAL PATHOGENS (Machine-translation by Google Translate, not legally binding) |
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